Sl/Sl4 hSCF220 cell line小鼠肝脏肥大细胞株 BioVector NTCC质粒载体菌种细胞基因保藏中心

Sl/Sl4 hSCF220 cell line小鼠肝脏肥大细胞株

Product category Animal cells Organism Mus musculus, mouse Classification Eukaryota, Animalia, Metazoa, Chordata, Vertebrata, Tetrapod Cell type mast cell Morphology fibroblast Tissue Liver General Specific applications SL/Sl4 hSCF220 is a transfected SL/Sl4 cell line that expresses membrane-bound stem cell factor (SCF). This cell line was derived from the Sl/Sl4 cell line (NTCC® CRL-2452) by transfection with spliced forms of hSCF cDNAs inserted into the pJT-1 (hSCF220) vector and expressed off the simian virus 40 (SV40) early promoter. Defects in the HM, associated with the Steel (Sl) mutation in mice, have been shown to be due to abnormalities in the production or presentation of the protein product of the Steel gene. Sl/Sl4 is a SV40 large T antigen immortalized stromal cell line derived from the hematopoietic microenvironment (HM) of a fetal murine homozygous (Sl/Sl) SCF-deficient embryo. It exists as a locally secreted or membrane-bound protein. SL/Sl4 hSCF220 cells do not secrete detectable amounts of SCF. Secreted SCF is available in the transfected Sl/Sl4 cell line, Sl/Sl4 hSCF248 (NTCC® CRL-2454). Characteristics Growth properties Adherent Derivation This cell line was derived from the Sl/Sl4 cell line (NTCC® CRL-2452) by transfection with spliced forms of hSCF cDNAs inserted into the pJT-1 (hSCF220) vector and expressed off the simian virus 40 (SV40) early promoter. Sl/Sl4 is a SV40 large T antigen immortalized stromal cell line derived from the hematopoietic microenvironment (HM) of a fetal murine homozygous (Sl/Sl) SCF-deficient embryo. Age embryo Comments SL/Sl4 hSCF220 is a transfected SL/Sl4 cell line that expresses membrane-bound stem cell factor (SCF). This cell line was derived from the Sl/Sl4 cell line (ATCC CRL-2452) by transfection with spliced forms of hSCF cDNAs inserted into the pJT-1 (hSCF220) vector and expressed off the simian virus 40 (SV40) early promoter. It was cotransfected with the p48 vector encoding hygromycin B using the Lipofectin method. Defects in the HM, associated with the Steel (Sl) mutation in mice, have been shown to be due to abnormalities in the production or presentation of the protein product of the Steel gene. Sl/Sl4 is a SV40 large T antigen immortalized stromal cell line derived from the hematopoietic microenvironment (HM) of a fetal murine homozygous (Sl/Sl) SCF-deficient embryo. This product is termed stem cell factor (SCF) or mast cell growth factor (MGF). It exists as a locally secreted or membrane-bound protein. Both forms show biological activity when assayed on human hematopoietic cells. SL/Sl4 hSCF220 cells that express the membrane-bound form (hSCF220) of SCF supported the maintenance of human progenitor cells in culture for up to four weeks This was 1-2 weeks longer than SL/Sl4 hSCF248. SL/Sl4 hSCF220 cells do not secrete detectable amounts of SCF. Secreted SCF is available in the transfected Sl/Sl4 cell line, Sl/Sl4 hSCF248 (ATCC CRL-2354). Handling information Unpacking and storage instructions Check all containers for leakage or breakage. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use. Complete medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: bovine calf serum to a final concentration of 10%. Temperature 37°C Atmosphere 95% Air, 5% CO2 Handling procedure To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at –70°C. Storage at –70°C will result in loss of viability. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes). Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. Transfer the vial contents to a 75 cm2 tissue culture flask coated with 0.1% gelatin and dilute with the recommended complete culture medium (see the specific batch information for the recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet. Note: If it is desired that the cryoprotective agent be removed immediately, or that a more concentrated cell suspension be obtained, centrifuge the cell suspension at approximately 125 x g for 5 to 10 minutes. Discard the supernatant and resuspend the cells with fresh growth medium at the dilution ratio recommended in the specific batch information. Subculturing procedure Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Subculture when flasks reach 80% to 90% confluency. Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels coated with 0.1% gelatin. Incubate cultures at 37°C. Subcultivation Ratio: 1:10 to 1:30 Medium Renewal: Every 2 to 3 days Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. Reagents for cryopreservation Complete growth medium supplemented with 5% (v/v) DMSO

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